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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
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Updated: Sep 21, 2025

Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Computationally designed hyperactive Cas9 enzymes.

Pascal D Vos1,2,3,4,5,6, Giulia Rossetti3,4,7, Jessica L Mantegna1,2,3,4

  • 1Curtin Medical School, Curtin University, Bentley, WA, Australia.

Nature Communications
|May 31, 2022
PubMed
Summary
This summary is machine-generated.

Researchers computationally designed hyperactive Cas9 enzymes using FuncLib, significantly improving gene editing efficiency. These advanced CRISPR-based tools enable larger and more diverse genomic modifications in mammalian cells.

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Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Last Updated: Sep 21, 2025

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Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Genetic engineering is crucial for understanding gene function and modifying organisms.
  • Current gene editing technologies face technical limitations, hindering their full potential.
  • CRISPR-Cas9 systems offer powerful gene editing capabilities but require optimization.

Purpose of the Study:

  • To computationally design Cas9 enzymes with enhanced donor-independent editing activities.
  • To develop improved tools for CRISPR-based gene editing applications.
  • To overcome existing technical challenges in genetic engineering.

Main Methods:

  • Utilized FuncLib for computational design of Cas9 enzymes.
  • Employed genetic circuits linked to cell survival in yeast for activity quantification.
  • Assessed engineered Cas9 variants in mammalian cells for editing efficiency and outcomes.

Main Results:

  • Designed and identified hyperactive Cas9 variants with substantially higher editing activity.
  • Discovered synergistic interactions between engineered regions in Cas9 enzymes.
  • Demonstrated efficient genome editing in mammalian cells with increased insertion and deletion diversity.

Conclusions:

  • FuncLib enables the design of highly active Cas9 enzymes, advancing gene editing.
  • Engineered Cas9 variants offer enhanced capabilities for CRISPR-based gene editing.
  • These tools expand the potential applications of genome modification in research and biotechnology.